This application is a 371 national stage application of PCT/JP97/03016, filed Aug. 29, 1997, which claims the benefit of priority to Japan Application No. 233316/96, filed Sep. 03, 1996.
The present invention relates to a method for inducing imniunosuppressive cells and a culture device to be used therefor. More particularly, the present invention relates to a method for inducing immunosuppressive cells by culturing human cells with the use of a culture device having an affinity for protein, and the culture device to be used therefor.
Autoimmune diseases and rejection in transplantation are caused by the following: host cells recognize self-cells or an organ transplanted as a foreign body and destroy the self-cells or the organ transplanted. Some cases of those diseases show that they are caused by abnormality of immunosuppressive cells in quantity or quality (see Macintosh and Drachman, Science vol. 232, p. 401(1986)). Balashov et al. report that MS (multiple sclerosis) patients show remarkable decrease in efficiency for inducing immunosuppressive T cells due to an autologous mixed lymphocyte reaction and such phenomenon is a cause of an attack of the MS disease (see Balashov et al., Journal of Clinical Investigation, vol. 95, p. 2711(1995)). Accordingly, it is considered that the recovery of the immunosuppressive cells of such patients may be effective in a therapeutic method for such patients. Thus for several examples of inducing immunosuppressive T cells in a system using animals such as mice are reported by, for example, Fresno et al. (see Fresno et al., Journal of Experimental Medicine, Vol. 163, p. 1246 (1980)). Further, as an example of inducing the immunosuppressive cells in human, there is a method utilizing the autologous mixed lymphocyte reaction described in the above reference which has been recently reported. However, this method requires the treatment of a culture medium of human cells with PHA, Con A or anti-CD3 antibody, or adding a plurality of cytokines to the culture. Thus, the method is complicated in operation. Moreover, since the above-mentioned method is suspension culture system, foreign bodies such as lectins (PHA or ConA) used in treatment adhere to the cells. Since those foreign bodies can not be removed completely from the cells by washing, the above-mentioned method still has the possibility of foreign body contamination. Accordingly, the method would never be considered to be desirable and thus, can not be utilized in treatment that targets induction of immunosuppressive cells outside of the body of a patient. The present invention provides an efficient therapeutic system for autoimmune diseases which includes the steps of: taking blood cells out from a patient; effectively inducing immunosuppressive cells, especially immunosuppressive T cells using an immobilized-protein system; and returning the resulting blood cells to the patient.
The present inventors have studied to improve a technique for preparing cells which selectively suppress hypersensitivity of the immune system which causes autoimmune diseases such as those mentioned above, especially immunosuppressive T cells and have completed the present invention.
The present invention provides a method for inducing immunosuppressive cells by culturing human cells with the use of (1) a culture device having an affinity for protein, being previously coated with a protein such as an antibody or a cytokine, or without any coating, or (2) a culture device having an affinity for protein, placing a protein such as the antibody or the cytokine in the device together with the human cells; and the culture device to be used therefor.
That is, the present invention relates to a method for inducing immunosuppressive cells, which comprises culturing human cells with the use of a culture device having an affinity for protein.
Further, the present invention relates to the inducing method, wherein the culture device is previously coated with one or more cytokines or antibodies against surface antigens (claim 2).
Moreover, the present invention relates to the inducing method of, wherein the culture device is previously coated with two or more antibodies against surface antigens, each of said antibodies recognizing different epitope (claim 3).
Further, the present invention relates to the inducing method, wherein human cells and one or more cytokines or antibodies against surface antigens are mixed.
Furthermore, the present invention relates to the inducing method, wherein human cells and two or more antibodies against surface antigens, each of said antibodies recognizing different epitope, are mixed (claim 5).
Moreover, the present invention relates to the inducing method, wherein the antibody against surface antigens is an anti-CD2 antibody or an anti-CD3 antibody.
Further, the present invention relates to the inducing method, wherein the antibody against surface antigens is an anti-CD2 antibody, said anti-CD2 antibody binding to a site of CD2 that participates in the binding of LFA-3 to CD2.
Further, the present invention relates to the inducing method, wherein the antibody against surface antigens is an anti-CD2 antibody, said anti-CD2 antibody binding to a site of CD2 other than a site of CD2 which participates in the binding of LFA-3 to CD2.
Moreover, the present invention relates to the inducing method, wherein two or more antibodies against surface antigens are anti-CD2 antibodies comprising a combination of one or more antibodies which bind to a site of CD2 that participates in the binding to LFA-3 to CD2 and one or more antibodies which bind to a site of CD2 other than the site of CD2 which participates in the binding of LFA-3 to CD2.
Further, the present invention relates to the inducing method, wherein one antibody against surface antigens is the anti-CD2 antibody TS2/18.
Moreover, the present invention relates to the inducing method, wherein one antibody against surface antigens is the F(ab)2 fragment of the anti-CD2 antibody TS2/18.
Further, the present invention relates to the inducing method, wherein the cytokine is IL-2 or GM-CSF.
Moreover, the present invention relates to the inducing method, wherein 0.5 to 10% by volume of serum based on a culture medium is mixed in the culture.
Further, the present invention relates to the inducing method, wherein no serum is mixed with the culture medium in the culture.
Moreover, the present invention relates to the inducing method, wherein a term of the culture ranges between 1 and 7 days.
Further, the present invention relates to the inducing method, wherein the culture device is made of material having an affinity for protein.
Moreover, the present invention relates to the inducing method, wherein culture device is made of plastic material.
Further, the present invention relates to the inducing method, wherein the culture device is made of glass material.
Moreover, the present invention relates to the inducing method, wherein the culture device is a closed plane plate vessel or a closed vessel charged with spherical fine-particles.
Further, the present invention relates to culture device having an affinity for protein, which is used for inducing immunosuppressive cells by culturing human cells.
Moreover, the present invention relates to the culture device, wherein the device is previously coated with one or more cytokines or antibodies against surface antigens.
Further, the present invention relates to the culture device, wherein the device is previously coated with two or more antibodies against surface antigens, each of said antibodies recognizing different epitope.
Moreover, the present invention relates to the culture device, wherein the antibody against surface antigens is anti-CD2 antibody or anti-CD3 antibody.
Further, the present invention relates to the culture device, wherein one or more antibodies that recognize different epitope are the anti-CD2 antibody TS2/18.